G418 Sulfate, also known as Geneticin Sulfate, is an amino-glycoside antibiotic and structurally similar to Gentamycin B1. It interfers with protein synthesis by binding 80s ribosome and blocking elongation steps. G418 Sulfate is toxic to both prokaryotic and eukaryotic cells, including bacteria, yeast, higher plant and mammalian cells, as well as protozoans and worms. The resistance gene (mainly neo), locating in transposon Tn601 (903) or Tn5, is derived from bacteria, but can be expressed in eukaryotic cells. The resistance gene can be introduced into cells through gene recombination techniques to obtain resistance of G418, which could be used to screen and maintain the culture of prokaryotic or eukaryotic cells carrying resistance gene.
In mammalian cells, neo,encodes the expression of amino-glycoside 3' -phosphotransferase (APH (3') II) after integrated into the eukaryotic genome. This enzyme inactivates the antibiotic by covalently modifying the amino or hydroxyl function of G418 and inhibiting the antibiotic-ribosome interaction, endowing the cell with G418 resistance. In screening stable cell line experiments, the killing curves (dose-response curves) should be established to determine the minimum effective concentration for killing non-resistant cells.
In plant cells, resistance can be obtained by transfection of nptII gene resistant plasmid. The nptII gene also encodes aminoglycoside phosphotransferase, an enzyme that inactivates several antibiotics, including G418, kanamycin, and palomycin.
Screening of stably transfected cell lines
|Synonym||G-418 disulfate; Geneticin sulfate; Antibiotic G418; G 418 Sulfate; Antibiotic G-418 sulfate|
|Molecular weight||692.7 g/mol|
|Appearance||White or light white powder|
|Potency（anhydrous）||> 700 U/mg|
|Solubility||≥60 mg/mL in H2O; insoluble in EtOH; insoluble in DMSO|
|60220||G418 Sulfate (Geneticin)||1 g||5 g|
The product should be stored at -25℃ ~ -15℃ for 3 years. Avoid moisture, which can reduce antibiotic activity.
Figure 1 Stability test and validation of effective concentration of genomycin between different batches
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